LOCUS       sequence-322782-        6802 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a TFR and ST6GAL1 swapped chimera with the cytosolic tail 
            and TMD from ST6GAL1, and luminal domain from TFR in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6802)
  AUTHORS   Sun X, Mahajan D, Chen B, Song Z, Lu L
  TITLE     A quantitative study of the Golgi retention of glycosyltransferases.
  JOURNAL   J Cell Sci. 2021 Oct 15;134(20). pii: 272560. doi: 
            10.1242/jcs.258564. Epub 2021 Oct 21.
  PUBMED    34533190
REFERENCE   2  (bases 1 to 6802)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2021-10-15"; volume: "134(20). pii: 272560. doi"; 
            pages: " 10.1242/jcs.258564. Epub 2021 Oct 21"
COMMENT     Sequence Label: SSF-GFP
FEATURES             Location/Qualifiers
     source          1..6802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1408..1419)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2764..2773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2770..2772,2773..2775,2776..3489)
                     /codon_start=1
                     /product="monomeric GFP variant optimized for use in 
                     oxidizing environments such as the secretory pathway 
                     (Costantini et al., 2015)"
                     /label=moxGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2815..2836)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3611..3732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3648..3667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3702..3721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3739..4194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4088..4107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4221..4325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4327..4684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4327..4518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4350..4370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4535..4670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4719..5513
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4719..5513
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4773..4792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5383..5402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5701..5720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5745..5792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6121..6709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6610..6629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagcgct accggactca gatctcgagg ccaccatgat tcacaccaac
      661 ctgaagaaaa agttcagctg ctgcgtcctg gtctttcttc tgtttgcagt catctgtgtg
      721 tggaaggaaa agtgtaaagg ggtagaacca aaaactgagt gtgagagact ggcaggaacc
      781 gagtctccag tgagggagga gccaggagag gacttccctg cagcacgtcg cttatattgg
      841 gatgacctga agagaaagtt gtcggagaaa ctggacagca cagacttcac cagcaccatc
      901 aagctgctga atgaaaattc atatgtccct cgtgaggctg gatctcaaaa agatgaaaat
      961 cttgcgttgt atgttgaaaa tcaatttcgt gaatttaaac tcagcaaagt ctggcgtgat
     1021 caacattttg ttaagattca ggtcaaagac agcgctcaaa actcggtgat catagttgat
     1081 aagaacggta gacttgttta cctggtggag aatcctgggg gttatgtggc gtatagtaag
     1141 gctgcaacag ttactggtaa actggtccat gctaattttg gtactaaaaa agattttgag
     1201 gatttataca ctcctgtgaa tggatctata gtgattgtca gagcagggaa aatcaccttt
     1261 gcagaaaagg ttgcaaatgc tgaaagctta aatgcaattg gtgtgttgat atacatggac
     1321 cagactaaat ttcccattgt taacgcagaa ctttcattct ttggacatgc tcatctgggg
     1381 acaggtgacc cttacacacc tggattccct tccttcaatc acactcagtt tccaccatct
     1441 cggtcatcag gattgcctaa tatacctgtc cagacaatct ccagagctgc tgcagaaaag
     1501 ctgtttggga atatggaagg agactgtccc tctgactgga aaacagactc tacatgtagg
     1561 atggtaacct cagaaagcaa gaatgtgaag ctcactgtga gcaatgtgct gaaagagata
     1621 aaaattctta acatctttgg agttattaaa ggctttgtag aaccagatca ctatgttgta
     1681 gttggggccc agagagatgc atggggccct ggagctgcaa aatccggtgt aggcacagct
     1741 ctcctattga aacttgccca gatgttctca gatatggtct taaaagatgg gtttcagccc
     1801 agcagaagca ttatctttgc cagttggagt gctggagact ttggatcggt tggtgccact
     1861 gaatggctag agggatacct ttcgtccctg catttaaagg ctttcactta tattaatctg
     1921 gataaagcgg ttcttggtac cagcaacttc aaggtttctg ccagcccact gttgtatacg
     1981 cttattgaga aaacaatgca aaatgtgaag catccggtta ctgggcaatt tctatatcag
     2041 gacagcaact gggccagcaa agttgagaaa ctcactttag acaatgctgc tttccctttc
     2101 cttgcatatt ctggaatccc agcagtttct ttctgttttt gcgaggacac agattatcct
     2161 tatttgggta ccaccatgga cacctataag gaactgattg agaggattcc tgagttgaac
     2221 aaagtggcac gagcagctgc agaggtcgct ggtcagttcg tgattaaact aacccatgat
     2281 gttgaattga acctggacta tgagaggtac aacagccaac tgctttcatt tgtgagggat
     2341 ctgaaccaat acagagcaga cataaaggaa atgggcctga gtttacagtg gctgtattct
     2401 gctcgtggag acttcttccg tgctacttcc agactaacaa cagatttcgg gaatgctgag
     2461 aaaacagaca gatttgtcat gaagaaactc aatgatcgtg tcatgagagt ggagtatcac
     2521 ttcctctctc cctacgtatc tccaaaagag tctcctttcc gacatgtctt ctggggctcc
     2581 ggctctcaca cgctgccagc tttactggag aacttgaaac tgcgtaaaca aaataacggt
     2641 gcttttaatg aaacgctgtt cagaaaccag ttggctctag ctacttggac tattcaggga
     2701 gctgcaaatg ccctctctgg tgacgtttgg gacattgaca atgagtttcg ggatccaccg
     2761 gtcgccacca tggtgtccaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2821 gagctggacg gcgacgtaaa cggccacaag ttctccgtgc ggggcgaggg cgagggcgat
     2881 gccaccaacg gcaagctgac cctgaagttc atcagcacca ccggcaagct gcccgtgccc
     2941 tggcccaccc tcgtgaccac cctgacctac ggcgtgcaga gcttctcccg ctaccccgac
     3001 cacatgaagc gccacgactt cttcaagagc gccatgcccg aaggctacgt ccaggagcgc
     3061 accatctcct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc
     3121 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3181 ctggggcaca agctggagta caacttcaac tcccacaacg tctatatcac cgccgacaag
     3241 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtggagga cggctccgtg
     3301 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3361 gacaaccact acctgtccac ccagtccaag ctgtccaaag accccaacga gaagcgcgat
     3421 cacatggtcc ttctggaatt cgtgaccgcc gccgggatca ctcacggcat ggacgagctg
     3481 tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     3541 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3601 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3721 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3781 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3841 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3901 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3961 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4021 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4081 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4141 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4261 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4321 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4381 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4621 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4681 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5221 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5701 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5761 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5821 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5881 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5941 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6241 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6781 gttatcccct gattctgtgg at
//
